1. Field of the Invention
The present invention relates to ionic complex nanoparticles, and particularly, to ionic complex nanoparticles for detecting heparanase activities and a method for preparing the same.
2. Background of the Invention
An extracellular matrix (ECM) serves to fill extracellular spaces of each organ and tissue, and constitutes a complex network of macromolecules for cells inside each organ. The main components of a basement membrane include collagen (type IV collagen), laminin, and heparan sulfate proteoglycan (HSPG). The HSPG is present on cell surfaces as well as in the ECM. And, the HSPG is composed of proteoglycan (coreprotein+glycosaminoglycan), and heparan sulfate (HS). The HSPG has a repetitive structure of disaccharide of N-acetylglucosamine and glucuronic acid. HS chains fill a main space of the basement membrane. The HS may bind to a variety of proteins or growth factors, and regulate functions of the proteins. And, the HS may serve as intracellular signaling pathways between growth factors and growth factor receptors.
Cells which are invading metastatic cancer cells or leukocytes or tissues of various inflammatory diseases pass through barriers of the extracellular matrix and the basement membrane with secreting protease. Here, the protease mainly belongs to a group consisting of matrix metalloprotease (MMP), serine, cysteine and aspartic protease, and serves to degrade the ECM. The heparanase is an endoglycosidase that specifically degrades the heparan sulfate, and is a degradation enzyme that plays an important role in invasions, metastases, and growths of cells. The heparanase plays a key role in invasion to cells, and also has biological activities such as angiogenesis by dissociating HS-growth factors from the ECM.
According to the recent researches, it was newly found that a degradation enzyme such as protease and heparanase plays an important role to cause various human disease such as cancers and dementia. Various recent research has reported that matrix metalloprotease (MMP) and heparanase serve to degrade the extracellular matrix in cells and in vivo, and are related to a cell mobility due to degradation of a pericellular matrix. And, it was investigated that the degradation enzymes play an important role in cancer growths such as angiogenesis, invasion of cancer cells and metastasis. Accordingly, there have been efforts among mega pharmaceutical companies to develop novel drugs with targeting the protease and heparanase.
The heparanase plays an important role in tumor cell invasion, metastasis, and various inflammatory diseases such as rheumatoid arthritis. However, due to the lack of research on a method for imaging and analyzing activities and expressions of the heparanase, or a method for non-invasively imaging an expressed degree of the heparanase in vivo, the related technologies are required.
The conventional methods for detecting the heparanase include 2D (two-dimensional) gel methods, multi-dimensional liquid chromatography methods. However, these methods require multi-step for measuring protocols, resulting in inefficiencies in the economic and time aspects when screening plenty of drugs in the process of developing novel drugs. For detection of heparanase activities, it has been developed a sensor using polymers and XL665-streptavidin conjugate, wherein the polymers are formed by binding Europium cryptate and Biotin to HSPG (see: K. Enomoto, H. Okamoto, Y Numata, H. Takemoto, J. Pharm. Biomed, Anal, 2006, 41m 912-917). This sensor has been used as a kit for detecting heparanase activities. However, it is difficult to detect the heparanase activities expressed in cells and in vivo, or to early diagnose diseases by using the sensor.